Abdolreza Ardeshirylajimi

Improved proliferation and osteogenic differentiation of mesenchymal stem cells on polyaniline composited by polyethersulfone nanofibers

Tissue engineering is a promising emerged method trying to reconstruct lost tissues that using synthetic and biomaterials and their combination with cells. The purpose of this study is increase osteoinductivity of polyethersulfone (PES) by using polyaniline (PANi). In this study, after fabrication of PES and composited PES-PANi scaffolds by electrospinning, scaffolds were characterized morphologically and mechanically. Then osteoinductivity of scaffolds was investigated by osteogenic differentiation of human mesenchymal stem cells (MSCs) cultured on the PES and PES-PANI in comparison to the tissue culture polystyrene as a control. The Osteogenic potential of MSCs was evaluated by Alizarin Red staining, ALP activity, calcium content assay and bone related gene expression assay. Scaffolds were smooth, bead free and in the scale of nanometers and PES mechanical stability was decreased significantly after composite with PANi. Highest growth, ALP activity and deposited calcium of cells were observed in the PES-PANi groups compared to the PES and TCPS. Significantly higher expression of collagen type one and osteonectin was also detected in the PES-PANi group. It can be concluded that PES-PANi construct has potential to be a good candidate as bone grafting substitute and using in tissue engineering applications.

Osteogenic differentiation Potential of Mesenchymal Stem Cells cultured on Nanofibrous Scaffold Improved in the Presence of Pulsed Electromagnetic Field

Nowadays, tissue engineering by using stem cells in combination with scaffolds and bioactive molecules has made significant contributions to the regeneration of damaged bone tissues. Since the usage of bioactive molecules including, growth factors to induce differentiation is safety limited in clinical applications, and it has also been previously observed that extremely low frequency pulsed electromagnetic fields (PEMF) can be effective in the enhancement of proliferation rate and osteogenic differentiation of stem cells, the aim of this study was investigating the osteoinductive potential of PEMF in combination with Poly(caprolactone) (PCL) nanofibrous scaffold. To achieve this aim, Adipose‐derived mesenchymal stem cells (ADSCs) isolated and characterized and then osteogenic differentiation of them was investigated after culturing on the surface of PCL scaffold under treatments of PEMF, PEMF plus osteogenic medium (OM) and OM. Analysis of common osteogenic markers such as Alizarin red staining, ALP activity, calcium content and four important bone‐related genes in days of 7, 14, and 21 confirmed that the effects of PEMF on the osteogenic differentiation of ADSCs are very similar to the effects of osteogenic medium. Thus, regarding the immunological concerns about the application of bioactive molecules for tissue engineering, PEMF could be a good alternative for osteogenic medium. Although, results were showed a synergetic effect for simultaneous application of PEMF and PCL scaffold in the osteogenesis process of ADSCs. Taking together, ADSCs‐seeded PCL nanofibrous scaffold in combination with PEMF could be a great option for use in bone tissue engineering applications.

Insulin producing cells generation by overexpression of miR-375 in adipose-derived mesenchymal stem cells from diabetic patients

Diabetes Mellitus (DM) is a systematic disease, which happens because of destruction of islets of Langerhans in the pancreas and systematic resistance to insulin. The lack of donor for pancreas transplantation and risk of transplant rejection is the main challenge in the treatment of this disease. Stem cells are proper and sufficient source for creating insulin-producing cells (IPC). In this study adipose tissue was provided from diabetic patients operated for liposuction and then adipose derived stem cells (ADSCs) were isolated, characterized and then treated by lentiviruses containing miR-375, after 7, 14 and 21 days of induction, islet-like clusters (ILC) specific genes including insulin and PDX1 were evaluated by Real Time RT-PCR. Finally, immunocytochemistry was also used for evaluation of these markers in the protein level. The results were shown that insulin and PDX1 genes and proteins expression significantly increased in transduced stem cells compared to the control group. According to the results it can be concluded that islet-like clusters can be achieved from ADSCs by overexpression of miR-375.

Improvement of hepatogenic differentiation of iPS cells on an aligned polyethersulfone compared to random nanofibers

Abstract The application of stem cells holds great promises in cell and tissue transplants. This study was designed to compare the hepatogenic differentiation of iPSCs on aligned PES/COL versus random. Aligned and random PES/COL nanofibrus scaffolds were fabricated by electrospining and their surface modified through plasma treatment and collagen coating. The scaffolds were characterized using scanning electron microscopy (SEM) and ATR-FTIR. Morphology and biochemical activities of the differentiated hepatocyte-like cells (HLCs) were examined after 5 and 20 days of differentiation. Real-Time RT-PCR and ICC showed no significant difference in the mRNA and protein levels of two important definitive endoderm specific markers, including Sox17 and Foxa2 between two scaffolds. However, Real-Time RT-PCR analysis indicated an increase in the expression of Cyp7A1 gene over the period of the differentiation procedure on the aligned nanofibers but there was no difference in other genes such as Albumin and CK19. Moreover, comparison of hepatogenic differentiation evaluated by Albumin production in conditioned media of HLCs differentiated on aligned PES/COL, showed increase expression of these markers after 20 days compared to that of the random nanofibers. Taken together, the results of this study may indicate that aligned PES/COL nanofibrous scaffolds can improve terminal differentiation of HLCs from iPSCs.

Applied Induced Pluripotent Stem Cells in Combination With Biomaterials in Bone Tissue Engineering: INDUCED PLURIPOTENT STEM CELLS'S OSTEOGENESIS

Due to increasing of the orthopedic lesions and fractures in the world and limitation of current treatment methods, researchers, and surgeons paid attention to the new treatment ways especially to tissue engineering and regenerative medicine. Innovation in stem cells and biomaterials accelerate during the last decade as two main important parts of the tissue engineering. Recently, induced pluripotent stem cells (iPSCs) introduced as cells with highly proliferation and differentiation potentials that hold great promising features for used in tissue engineering and regenerative medicine. As another main part of tissue engineering, synthetic, and natural polymers have been shown daily grow up in number to increase and improve the grade of biopolymers that could be used as scaffold with or without stem cells for implantation. One of the developed areas of tissue engineering is bone tissue engineering; the aim of this review is present studies were done in the field of bone tissue engineering while used iPSCs in combination with natural and synthetic biomaterials. J. Cell. Biochem. 118: 3034–3042, 2017.

Adapted dexamethasone delivery polyethylene oxide and poly(ɛ-caprolactone) construct promote mesenchymal stem cells chondrogenesis

Abstract Issues associated with tissue transplantation and shortage of donors has always been a concern, whereas tissue engineering has provided the hopeful opportunities. The aim of this study was to investigate the chondrogenic differentiation potential of mesenchymal stem cells (MSCs) in the presence of embedded Dexamethasone into electrospun Poly(ethyleneoxide) nanofibers composited with Poly(ɛ-caprolactone) nanofibers. Electrospun-fabricated scaffolds were characterized by SEM, tensile, contact angle, release profile, MTT assay, and chonderogenic differentiation of stem cells. Results demonstrated Poly(ɛ-caprolactone) properties improved by composite with a Poly(ethyleneoxide). Overall, stem cells seeded on nanofibers content drug showed highest chondrogenic potential in comparison to the other groups.

Prolonged drug release using PCL–TMZ nanofibers induce the apoptotic behavior of U87 glioma cells

ABSTRACT In situ prolonged delivery of drugs at the site of tumor can be satisfactorily accelerated patient recovery. We compared the effect of temozolomide while incorporated by polycaprolactone nanofibers on the apoptotic behavior of U87 glioma cells. After biocompatibility evaluation of nanofibers by scanning electron microscope and 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide analysis, the apoptosis of U87 cells was evaluated using p53, Bcl2 and Bax genes expression. It was found that nanofiber-temozolomide group showed a greater ability to induce apoptosis as well as have a significantly diminished initial burst release of drug compared with other groups and have promising potential in treating cancer. GRAPHICAL ABSTRACT

Bioactive glass ceramic nanoparticles-coated poly(l-lactic acid) scaffold improved osteogenic differentiation of adipose stem cells in equine

Horses with big bone fractures have low chance to live mainly due to the lake of a proper treatment strategy. We believe that further attempts in equine bone tissue engineering will probably be required to meet all the needs for the lesion therapies. Therefore in this study we aimed to investigate the osteogenic differentiation capacity of equine adipose-derived stem cells (e-ASCs) on nano-bioactive glass (nBGs) coated poly(l-lactic acid) (PLLA) nanofibers scaffold (nBG-PLLA). Using electrospinning technique, PLLA scaffold was prepared successfully and coated with nBGs. Fabricated nanofibers were characterized by MTT, SEM, and FTIR analyses, and then osteogenic differentiation potential of isolated e-ASCs was investigated by the most key osteogenic markers, namely Alizarin red-S, ALP, calcium content and bone related (RUNX2, Collagen I, Osteonectin, and ALP) gene markers. Our results indicated that nBGs was successfully coated on PLLA scaffold and this scaffold had no negative (p>0.05) effect on cell growth rate as indicated by MTT assay. Moreover, e-ASCs that differentiated on nBGs-PLLA scaffold showed a higher (p<0.05) ALP activity, more (p<0.05) calcium content, and higher (p<0.05) expression of bone-related genes than that on uncoated PLLA scaffold and TCPS. According to the results, a combination of bioceramics and biopolymeric nanofibers hold valuable promising potentials to use for bone tissue engineering application and regenerative medicine.

Evaluation of hypoxia inducible factor-1 alpha gene expression in colorectal cancer stages of Iranian patients

AIM OF STUDY Colorectal cancer (CRC) is the fourth most prevalent cancer globally. Several factors have roles in cancer establishment. One of the most important factors is hypoxia that induces hypoxia inducible factor-1 (HIF-1). The HIF-1 alpha overexpressed in hypoxia conditions and plays a pivotal role in carcinogenesis features. In this study, we aimed to examine the efficiency of HIF-1 alpha gene expression at mRNA and proteins level for CRC diagnosing and staging. MATERIALS AND METHODS In this study, the cases included into 75 cancer specimens in different stages (Group 2 = Stage 1, Group 3 = Stage 2, and Group 4 = Stage 3, 4) and ten normal specimens as control (Group 1). Real-time reverse transcription-polymerase chain reaction and immunohistochemistry (IHC) were performed for measuring gene expression at RNA and proteins level, respectively. The raw data were analyzed in the SPSS20 software. RESULTS HIF-1 alpha gene expression rate (2-ΔΔCT) and ΔCT values were significantly higher increased in Group 4 in compare to control (P < 0.001). Other cancer groups (2 and 3) had greater ΔCT values than control, but it was not statistically significant. Moreover, the rate of HIF-1 alpha gene expression (2-ΔΔCT) was increased with cancer stages. According to the IHC results, there was a positive relationship between CRC stages and HIF-1 alpha protein expression (P < 0.05). CONCLUSIONS HIF-1 alpha gene expression increased in earlier up to metastasis stages of CRC, but the assessment of HIF-1 alpha gene expression has not important role in the diagnosis of cancer in early stages and classification of carcinoma because the increasing of HIF-1 alpha gene expression is not significant in early cancer stages.

Improved anticancer properties of stem cells derived exosomes by prolonged release from PCL nanofibrous structure

Systemic chemotherapy as a routine method for the treatment of cancers has several complications. Localized chemotherapy can significantly increase the treatment efficacy and reduce side effects. Here, mesenchymal stem cells derived microvesicles (MVs) was incorporated into Polycaprolactone nanofibers and then the apoptotic behavior of MCF-7 breast cancer cells was investigated while cultured on MVs-Polycaprolactone by using SEM, MTT and apoptotic gene expression. Results demonstrated that MVs-Polycaprolactone group was shown a great apoptosis induction potential into MCF-7 cells and it can be concluded that MVs-Polycaprolactone has a great promising potential to introduce as an in-situ tumor inhibitor construct rather than chemo-drugs.